Transcript Detection of human telomerase gene (TERC) amplification in

Detection of human telomerase
gene (TERC) amplification in
cervical neoplasia:
A retrospective study of 79 patients Pap
smear slides
Renarta Crookes
Sheffield Children's NHS Foundation Trust
Cervical Cancer



Papanicolou test reduced mortality rate
40,000 cases of mild/moderate
dyskaryosis sent for further investigation
HPV 16 or 18 accounts for 70% of cervical
cancer, majority of mild regress
CGH – Amplification of TERC
 CGH – 85% 3q26 TERC
3q26


The RNA component of
the human telomerase
3q gain in interphase
FISH as an indicator
of disease progression?
Categorisation of Smears


Negative
Low Grade


Mild dyskaryosis means the outer 1/3 of the
cervix is affected
High Grade


Moderate dyskaryosis means up to 2/3 of the
skin thickness is affected
CIN3 means the full thickness of the cervical
skin is affected
TERC – A new diagnostic marker?
 TERC gain is required for,
and predicts, progression
from mild/moderate
dyskaryosis to CIN3
 TERC gain observed in 33%
of negative Pap smears that
progressed to CIN3
100% of mild/moderate
cases that regressed did not
show TERC gains
Heselmeyer-Haddad et al., 2005. Am J. Pathol. 166 1229-1238
The Present Study

Aim – Can FISH for TERC
copy number be used to
improve the current
system for cervical smear
screening?
 79 retrospective cases
TERC 3q26
C-MYC 8q24
 Negative, mild or moderate progressing
to CIN3 or cervical cancer
Negative, mild or moderate regressing or
remaining negative
Results- Negative progressing to
CIN3
Increase in TERC copy number
Normal diploid cell
x100
x40
Results- Negative progressing to
CIN3
100
90
% of nuceli with
more than two
copies of TERC
80
70
60
50
40
% of nuclei with
more than two
copies of C-MYC
30
20
10
0
A
B
C
D
E
F
G
H
Patient
I
J
K
L
M
N
Results- Mild progressing to
CIN3
Increase in
C-MYC
Increase in TERC
Results- Mild progressing to
CIN3
100
% of nuclei
with more
than two
copies of
TERC
90
80
70
60
50
40
% of nuclei
with more
than two
copies of CMYC
30
20
10
0
A1
B1
C1
D1
E1
F1
G1
H1
Patient
I1
J1
K1
L1
M1
N1
O1
Results- Moderate progressing
to CIN3
82.4%
showed
70.6%
showed
moderate cases
gain of TERC
moderate cases
gain of C-MYC
Results- Moderate progressing
to CIN3
100
% of nuclei
with more
than two
copies of
TERC
90
80
70
60
50
% of nuclei
with more
than two
copies of
C-MYC
40
30
20
10
0
A2
B2
C2
D2
E2
F2
G2
H2
I2
P a t i e nt
J2
K2
L2
M2
N2
O2
P2
Q2
Summary of Progressors
100
90
% of cases
with more
than two
copies of
TERC
80
70
60
50
40
30
% of cases
with more
than two
copies of CMYC
20
10
0
Negative
Mild Dyskaryosis
Moderate
Dyskaryosis
Results- Negative remaining
Negative
100
90
80
% of nuclei with
more than two
copies of TERC
70
60
50
40
% of nuclei with
more than two
copies of C-MYC
30
20
10
0
A
B
C
D
E
F
G
H
Patient
I
J
K
L
M
N
O
Results- Mild regressing to negative
100
90
80
% of nuclei with
more that 2
copies of TERC
70
60
50
40
30
20
% of nuclei with
more than 2
copies of C-MYC
10
0
A1
B1
C1
D1
E1
F1
Patient
G1
H1
I1
J1
Results- Moderate regressing to
negative
100
90
80
% of nuclei with
more than 2
copies of TERC
70
60
50
40
% of nuclei with
more than 2
copies of C-MYC
30
20
10
0
A2
B2
C2
D2
E2
Patient
F2
G2
H2
Regressors
Negative and mild dyskaryosis
patients that regressed or
remained negative showed no
gains in TERC
2/8 patients classed as
moderate dyskaryosis
showed gains of TERC,
with a high proportion of
tetraploid cells.
Summary of Regressors
100
% of cases with
more than two
copies of TERC
90
80
70
60
% of cases with
more than two
copies of C-MYC
50
40
30
20
10
0
Negative
Mild Dyskaryosis
Moderate Dyskaryosis
Study Summary


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90 cases received
79 successful analyses
2 insufficient sample to score (min no.
cells to score?)
5 fails in progressors (thick smears)
4 fails in regressors (from 2000 - ?bond)
Application as a service



Analysis of slides ranges from 30 mins to over
an hour by eye per analyser
Automation – Scanning, Capture + Relocation
Liquid based cytology slides – reducing cell
clumping/false positive rate, no de-staining
Conclusions

TERC increases with severity of disease

Both negative and mild dyskaryosis
regressors showed no gains in TERC
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Prospective work using liquid based cytology
slides
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Trial automation to reduce analysis

Improve detection rate and reduce costs and
patient anxiety
Acknowledgments

Dr John H F Smith

Michael Dyson

Dr Edna L. Maltby

Abbott Vysis
Royal Hallamshire Hospital)
Service)
(Dept. of Histopathology,
(Sheffield Cytogenetics Service)
(Sheffield Cytogenetics